Substrate recognition induces sequential electron transfer across subunits in the nitrogenase-like DPOR complex
Abstract
A key step in bacteriochlorophyll biosynthesis is the reduction of protochlorophyllide (Pchlide) to chlorophyllide (Chlide), catalyzed by dark-operative protochlorophyllide oxidoreductase (DPOR). DPOR is made of electron donor (BchL) and acceptor (BchNB) component proteins. BchNB is further composed of two subunits each of BchN and BchB arranged as an α2β2 heterotetramer with two active sites for substrate reduction. Such oligomeric architectures are found in several other electron transfer (ET) complexes, but how this architecture influences activity is unclear. In this work, we describe allosteric communication between the two identical active sites in Rhodobacter sphaeroides BchNB that drives sequential and asymmetric ET. Pchlide binding to one BchNB active site initiates ET from the pre-reduced [4Fe-4S] cluster of BchNB, a process similar to the deficit spending mechanism observed in the structurally related nitrogenase complex. Pchlide binding in one active site is recognized in trans by an Asp-274 from the opposing half, which is positioned to serve as the initial proton donor. A D274A variant DPOR binds to two Pchlide molecules in the BchNB complex, but only one is bound productively, stalling Pchlide reduction in both active sites. A half-active complex combining one WT and one D274A monomer also stalled after one electron wasmore »
- Publication Date:
- Research Org.:
- Marquette Univ., Milwaukee, WI (United States)
- Sponsoring Org.:
- USDOE Office of Science (SC), Basic Energy Sciences (BES); National Science Foundation (NSF)
- OSTI Identifier:
- 1772158
- Alternate Identifier(s):
- OSTI ID: 1706081
- Grant/Contract Number:
- SC0017866; CHE-1532168
- Resource Type:
- Published Article
- Journal Name:
- Journal of Biological Chemistry
- Additional Journal Information:
- Journal Name: Journal of Biological Chemistry Journal Volume: 295 Journal Issue: 39; Journal ID: ISSN 0021-9258
- Publisher:
- Elsevier
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Electron transfer; chlorophyll; photosynthesis; metalloenzyme; EPR; electron transfer; nitrogenase; electron paramagnetic resonance; ATPase; cyanobacteria; DPOR; protochlorophyllide
Citation Formats
Corless, Elliot I., Bennett, Brian, and Antony, Edwin. Substrate recognition induces sequential electron transfer across subunits in the nitrogenase-like DPOR complex. United States: N. p., 2020.
Web. doi:10.1074/jbc.RA120.015151.
Corless, Elliot I., Bennett, Brian, & Antony, Edwin. Substrate recognition induces sequential electron transfer across subunits in the nitrogenase-like DPOR complex. United States. https://doi.org/10.1074/jbc.RA120.015151
Corless, Elliot I., Bennett, Brian, and Antony, Edwin. Tue .
"Substrate recognition induces sequential electron transfer across subunits in the nitrogenase-like DPOR complex". United States. https://doi.org/10.1074/jbc.RA120.015151.
@article{osti_1772158,
title = {Substrate recognition induces sequential electron transfer across subunits in the nitrogenase-like DPOR complex},
author = {Corless, Elliot I. and Bennett, Brian and Antony, Edwin},
abstractNote = {A key step in bacteriochlorophyll biosynthesis is the reduction of protochlorophyllide (Pchlide) to chlorophyllide (Chlide), catalyzed by dark-operative protochlorophyllide oxidoreductase (DPOR). DPOR is made of electron donor (BchL) and acceptor (BchNB) component proteins. BchNB is further composed of two subunits each of BchN and BchB arranged as an α2β2 heterotetramer with two active sites for substrate reduction. Such oligomeric architectures are found in several other electron transfer (ET) complexes, but how this architecture influences activity is unclear. In this work, we describe allosteric communication between the two identical active sites in Rhodobacter sphaeroides BchNB that drives sequential and asymmetric ET. Pchlide binding to one BchNB active site initiates ET from the pre-reduced [4Fe-4S] cluster of BchNB, a process similar to the deficit spending mechanism observed in the structurally related nitrogenase complex. Pchlide binding in one active site is recognized in trans by an Asp-274 from the opposing half, which is positioned to serve as the initial proton donor. A D274A variant DPOR binds to two Pchlide molecules in the BchNB complex, but only one is bound productively, stalling Pchlide reduction in both active sites. A half-active complex combining one WT and one D274A monomer also stalled after one electron was transferred in the WT half. We propose that such sequential electron transfer in oligomeric enzymes serves as a regulatory mechanism to ensure binding and recognition of the correct substrate. Overall, the findings shed light on the functional advantages imparted by the oligomeric architecture found in many electron transfer enzymes.},
doi = {10.1074/jbc.RA120.015151},
journal = {Journal of Biological Chemistry},
number = 39,
volume = 295,
place = {United States},
year = {Tue Sep 01 00:00:00 EDT 2020},
month = {Tue Sep 01 00:00:00 EDT 2020}
}
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